Correlation between longitudinal, circumferential, and radial moduli in cortical bone: effect of mineral content.
Previous studies indicate that changes in the longitudinal elastic properties of bone due to changes in mineral content are related to the longitudinal strength of bone tissue. Changes in mineral content are expected to affect bone tissue mechanical properties along all directions, albeit to different extents. However, changes in tissue mechanical properties along the different directions are expected to be correlated to one another. In this study, we investigate if radial, circumferential, and longitudinal moduli are related in bone tissue with varying mineral content. Plexiform bovine femoral bone samples were treated in fluoride ion solutions for a period of 3 and 12 days to obtain bones with 20% and 32% lower effective mineral contents. Transmission ultrasound velocities were obtained in the radial, circumferential, and longitudinal axes of bone and combined with measured densities to obtain corresponding tensorial moduli. Results indicate that moduli decreased with fluoride ion treatments and were significantly correlated to one another (r(2) radial vs. longitudinal = 0.80, r(2) circumferential vs. longitudinal = 0.90, r(2) radial vs. circumferential = 0.85). Densities calculated from using ultrasound parameters, acoustic impedance and transmission velocities, were moderately correlated to those measured by the Archimedes principle (r(2)=0.54, p<0.01). These results suggest that radial and circumferential ultrasound measurements could be used to determine the longitudinal properties of bone and that ultrasound may not be able to predict in vitro densities of bones containing unbonded mineral.


Prevention of worsening of severe thrombocytopenia after red cell transfusions by the use of leucocyte-depleted blood.
Platelet counts were measured before and after red cell transfusions in 30 patients with anaemia and severe thrombocytopenia resulting from haematological diseases. There was a mean reduction of 1.1 x 109/l (P = 0.43) in the platelet count after transfusions of 2-3 units of leucocyte-depleted red cell concentrates (20 patients). However, there was a mean reduction of 2.7 x 109/l (P = 0.03), approximately 10%, in the platelet count after transfusions of non-leucocyte-depleted red cell concentrates (10 patients). The findings suggest that the forthcoming introduction of universal leucocyte depletion of red cell concentrates will minimize the worsening of thrombocytopenia that occurs in severely thrombocytopenic patients receiving standard non-leucocyte-depleted red cell concentrates.


Anchored phylogenomics improves the resolution of evolutionary relationships in the rapid radiation of Protea L.
Estimating phylogenetic relationships in relatively recent evolutionary radiations is challenging, especially if short branches associated with recent divergence result in multiple gene tree histories. We combine anchored enrichment next-generation sequencing with species tree analyses to produce a robust estimate of phylogenetic relationships in the genus Protea (Proteaceae), an iconic radiation in South Africa. We sampled multiple individuals within 59 out of 112 species of Protea and 6 outgroup species for a total of 163 individuals, and obtained sequences for 498 low-copy, orthologous nuclear loci using anchored phylogenomics. We compare several approaches for building species trees, and explore gene tree-species tree discrepancies to determine whether poor phylogenetic resolution reflects a lack of informative sites, incomplete lineage sorting, or hybridization. Phylogenetic estimates from species tree approaches are similar to one another and recover previously well-supported clades within Protea, in addition to providing well-supported phylogenetic hypotheses for previously poorly resolved intrageneric relationships. Individual gene trees are markedly different from one another and from species trees. Nonetheless, analyses indicate that differences among gene trees occur primarily concerning clades supported by short branches. Species tree methods using hundreds of nuclear loci provided strong support for many previously unresolved relationships in the radiation of the genus Protea. In cases where support for particular relationships remains low, these appear to arise from few informative sites and lack of information rather than strongly supported disagreement among gene trees.


STRONTIUM 90: ESTIMATION OF WORLDWIDE DEPOSITION.
The relation between the worldwide deposition of strontium-90, as calculated by many investigators over the last decade, and that observed in rainfall in New York City has been relatively constant. On the average, for each millicurie of strontium-90 per square mile deposited in New York City, 0.055 megacurie has been deposited on the earth's total surface. Cumulative deposits of strontium-90 on the earth's surface at various intervals over the last 10 years have been computed from this ratio. From the mean quarterly fraction of the annual strontium-90 fallout in New York City for the last 9 years, the worldwide deposition of this nuclide, equal to 2.48 megacuries, is predicted for 1964.


Total laparoscopic donor nephrectomy in situs inversus totalis: a case report.
Situs inversus totalis is a rare anomaly characterized by the total inversion of all abdominal and thoracic organs. For the first time, we report a case of a donor nephrectomy in a patient with situs inversus totalis, completed with a full laparoscopic approach. At the time of this writing, the donor and the recipient are doing well after 6 month's follow-up. Our experience shows that patients with situs inversus totalis may be eligible candidates for laparoscopic donor nephrectomy, provided that detailed preoperative imaging studies as well as precise preoperative planning are performed before the transplant.


Comparison of fixed percentage method and lower confidence limits for defining limits of normality for interpretation of spirometry.
The use of the lower 90% confidence limit of the lower limit of normal (LLN(CI)), rather than a fixed percentage of the predicted value (LLN(%)), appears to be statistically more appropriate for interpretation of spirometry results. There has been no comparative assessment of these 2 definitions of the LLN in routine clinical practice. We studied results of spirometry interpretations made with these 2 approaches, and assessed various factors that influence discordant classification of spirometry results. Spirometry records from 18,112 consecutive adult patients referred for spirometry were interpreted as normal, obstructive, or restrictive, based on both LLN(CI) and LLN(%). Discordant results were analyzed using multiple logistic regression techniques to identify variables that significantly affected discordant classification of results. Overall, 11.7% of the results were discordant between the 2 methods. Agreement between the 2 methods, calculated using the kappa estimate, was poorer with spirometry values from women and from patients at the extremes of height and age. Age, sex, and height independently influenced discordant classification. Limits of agreement between LLN(CI) and LLN(%) were wide for all the spirometric variables studied--more so in women and in shorter and older patients. LLN(CI) and LLN(%) yielded different interpretations of spirometry data in several instances, and the 2 methods cannot be used interchangeably. When interpreting spirometry data in routine clinical practice, LLN(CI) should be preferred over LLN(%).


Functional Outcomes Following Laparoscopic Ileal Pouch-Anal Anastomosis in Patients with Chronic Ulcerative Colitis: Long-Term Follow-up of a Case-Matched Study.
Laparoscopic ileal pouch-anal anastomosis (L-IPAA) has been increasingly adopted over the last decade due to short-term patient-related benefits. Several studies have shown L-IPAA to be equivalent to open IPAA in terms of safety and short-term outcomes. However, few L-IPAA studies have examined long-term functional outcomes. We aimed to evaluate the long-term functional outcomes of L-IPAA as compared to open IPAA. A previous case-matched cohort study at our institution compared short-term outcomes between L-IPAA and open IPAA from 1998 to 2004. For this study, we selected all patients from this case-matched cohort study with chronic ulcerative colitis (CUC) who had follow-up functional data of greater than 1 year. Functional data was obtained through prospective surveys, which were sent annually to all IPAA patients postoperatively. One hundred and forty-nine patients (58 L-IPAA, 91 open IPAA) with a median 8-year duration of follow-up were identified. There were no differences in demographics and long-term surgical outcomes between groups. Stapled anastomosis was more common in the laparoscopic group (91.4 versus 54.9%, p < 0.001). Stool frequency during daytime (>6 stools, L-IPAA 32.8%, open 49.4%, p = 0.048) and nighttime (>2 stools, L-IPAA 13.8%, open 30.6%; p = 0.024) was significantly lower in the L-IPAA group. Ability to differentiate gas from stool was not different (p = 0.13). Rate of complete continence was similar in L-IPAA and open groups (L-IPAA 36.2%, open 21.8%, p = 0.060). There was no difference in use of medication to control stools, perianal skin irritation, voiding difficulty, sexual problems, and occupational change between groups. Subgroup analysis to evaluate for any group differences attributable to anastomotic technique demonstrated only that stapled anastomoses lead to more perianal skin irritation in the L-IPAA group (L-IPAA = 60.4% versus open IPAA = 38.8%; p = 0.031). Overall, L-IPAA has comparable functional results to the open approach with slightly lower daytime and nighttime stool frequency. This difference may be attributed to a greater number of stapled anastomoses performed in the laparoscopic cohort.


Stiff person syndrome in South Asia.
Stiff person syndrome is a highly disabling, progressive autoimmune disorder of the central nervous system characterized by muscle rigidity and spasms. Stiff person syndrome is rare, but is believed to be under diagnosed with only 14 cases been reported among a 1.7 billion population in South Asia. We report the first authenticated case from Sri Lanka. A 55-year-old Sri Lankan female presented with difficulty in walking and recurrent falls due to progressive muscular rigidity in her lower limbs and trunk with superimposed muscle spasms that occurred in response to unexpected noise, startle or emotional upset. She had anxiety and specific phobias to open spaces, walking unaided and being among crowds of people. She had insulin-dependent diabetes mellitus and was on thyroxine replacement. On examination, she had hyperlordosis combined with board-like rigidity of her anterior abdomen and rigidity of her lower limbs bilaterally. Upper limbs were normal. Magnetic resonance imaging of her neuraxis was normal. Electromyography showed continuous motor unit activity at rest. Glutamic acid decarboxylase antibodies were detected in her serum at a titre of 15,500 IU/ml (normal <5). She showed a remarkable and sustained improvement to treatment with intravenous immunoglobulins, immunosuppressive and muscle relaxant medications, regaining independent ambulation. Diagnosis of stiff person syndrome remains clinical, supported by electromyography and serology for glutamic acid decarboxylase antibodies, facilitated by a high index of clinical suspicion. An autoimmune basis lends stiff person syndrome amenable to treatment highlighting the importance of diagnosis. This case adds to map the worldwide distribution of stiff person syndrome.


PET/CT with (124)I-cG250: great potential and some open questions.
A novel PET radiotracer, (124)I-cG250, is currently under clinical investigation to distinguish clear cell renal cell carcinoma from other benign and malignant renal masses. In this article, we will make suggestions on the data needed to maximize the use of this radiotracer. Although the published data are promising, further data are needed to assess the potential usefulness of this agent when dealing with indeterminate renal masses.


The effective mutation rate at Y chromosome short tandem repeats, with application to human population-divergence time.
We estimate an effective mutation rate at an average Y chromosome short-tandem repeat locus as 6.9x10-4 per 25 years, with a standard deviation across loci of 5.7x10-4, using data on microsatellite variation within Y chromosome haplogroups defined by unique-event polymorphisms in populations with documented short-term histories, as well as comparative data on worldwide populations at both the Y chromosome and various autosomal loci. This value is used to estimate the times of the African Bantu expansion, the divergence of Polynesian populations (the Maoris, Cook Islanders, and Samoans), and the origin of Gypsy populations from Bulgaria.


Gender-specific modifying effect on the educational disparities in the impact of smoking on health expectancy.
Smoking reduces life years in good health but it is unclear how education modifies the impact of smoking. We hypothesize that the vulnerability of the effect of smoking on health expectancy decreases with educational level in both genders and examine the contributions of mortality and health effects. Life tables by educational level and smoking category were constructed from registers and survey data. For each educational level, difference in expected lifetime in self-rated good and poor health between 30-year-old never smokers and smokers were estimated and decomposed into contributions from mortality and health status. Difference in expected lifetime in good health between never smokers and smokers decreased with educational level for women but increased for men. Thus, the differences between never smokers and heavy smokers among 30-year-old women with a low, medium and high educational level were 12.9, 8.9 and 4.1 years, respectively. In contrast, the differences between male never smokers and heavy smokers with a low, medium and high educational level were 10.3, 11.4 and 14.3 years, respectively. Regardless of educational level, the mortality effect increased by exposure to smoking but the effect of health status increased by educational level for men and decreased for women. The social differential vulnerability to the effect of smoking differed between genders. Thus, whereas smoking had a substantial effect on health among women with a low educational level the pattern for men was opposite because the health gain for never smokers was greatest for men with a high education.


Comparison of plasmid- and chromosome-based polymerase chain reaction assays for detecting Chlamydia trachomatis nucleic acids.
Several laboratories have demonstrated that the polymerase chain reaction (PCR) is more sensitive than culture or enzyme immunoassay (EIA) for detecting Chlamydia trachomatis in genitourinary tract specimens when various DNA targets are used for amplification, including the cryptic plasmid, major outer membrane protein (MOMP), or rRNA genes. We compared the performances of five different PCR assays, including assays with two plasmid, two MOMP, and one rRNA targets, by amplifying serial dilutions of C. trachomatis DNA and testing genitourinary tract specimens. By using published procedures, two different plasmid primers had sensitivities of 0.1 fg for C. trachomatis plasmid DNA and 10 fg for total cellular DNA. The sensitivities of the assays with the two MOMP primers were 0.1 and 10 pg, and the sensitivity for the assay with the rRNA primers was 1 pg for cellular DNA. Both plasmid-based assays detected 38 of 38 confirmed Chlamydiazyme-positive specimens, whereas the assays with the MOMP and rRNA primers detected 36 of 38 and 29 of 38 confirmed Chlamydiazyme-positive specimens, respectively. Six of 18 Chlamydiazyme-negative specimens collected from individuals whose specimens were positive by culture or immunofluorescence were positive by both plasmid-based PCRs; 4 of these were positive by PCR with the MOMP primers and 3 were positive by PCR with the rRNA primers. The results obtained with both purified DNA and genitourinary tract specimens indicated that the plasmid-based PCRs are more sensitive than bacterial chromosome-based PCRs for detecting C. trachomatis.


European Society of Cardiology: Cardiovascular Disease Statistics 2019.
The 2019 report from the European Society of Cardiology (ESC) Atlas provides a contemporary analysis of cardiovascular disease (CVD) statistics across 56 member countries, with particular emphasis on international inequalities in disease burden and healthcare delivery together with estimates of progress towards meeting 2025 World Health Organization (WHO) non-communicable disease targets. In this report, contemporary CVD statistics are presented for member countries of the ESC. The statistics are drawn from the ESC Atlas which is a repository of CVD data from a variety of sources including the WHO, the Institute for Health Metrics and Evaluation, and the World Bank. The Atlas also includes novel ESC sponsored data on human and capital infrastructure and cardiovascular healthcare delivery obtained by annual survey of the national societies of ESC member countries. Across ESC member countries, the prevalence of obesity (body mass index ≥30 kg/m2) and diabetes has increased two- to three-fold during the last 30 years making the WHO 2025 target to halt rises in these risk factors unlikely to be achieved. More encouraging have been variable declines in hypertension, smoking, and alcohol consumption but on current trends only the reduction in smoking from 28% to 21% during the last 20 years appears sufficient for the WHO target to be achieved. The median age-standardized prevalence of major risk factors was higher in middle-income compared with high-income ESC member countries for hypertension {23.8% [interquartile range (IQR) 22.5-23.1%] vs. 15.7% (IQR 14.5-21.1%)}, diabetes [7.7% (IQR 7.1-10.1%) vs. 5.6% (IQR 4.8-7.0%)], and among males smoking [43.8% (IQR 37.4-48.0%) vs. 26.0% (IQR 20.9-31.7%)] although among females smoking was less common in middle-income countries [8.7% (IQR 3.0-10.8) vs. 16.7% (IQR 13.9-19.7%)]. There were associated inequalities in disease burden with disability-adjusted life years per 100 000 people due to CVD over three times as high in middle-income [7160 (IQR 5655-8115)] compared with high-income [2235 (IQR 1896-3602)] countries. Cardiovascular disease mortality was also higher in middle-income countries where it accounted for a greater proportion of potential years of life lost compared with high-income countries in both females (43% vs. 28%) and males (39% vs. 28%). Despite the inequalities in disease burden across ESC member countries, survey data from the National Cardiac Societies of the ESC showed that middle-income member countries remain severely under-resourced compared with high-income countries in terms of cardiological person-power and technological infrastructure. Under-resourcing in middle-income countries is associated with a severe procedural deficit compared with high-income countries in terms of coronary intervention, device implantation and cardiac surgical procedures. A seemingly inexorable rise in the prevalence of obesity and diabetes currently provides the greatest challenge to achieving further reductions in CVD burden across ESC member countries. Additional challenges are provided by inequalities in disease burden that now require intensification of policy initiatives in order to reduce population risk and prioritize cardiovascular healthcare delivery, particularly in the middle-income countries of the ESC where need is greatest.


Consensus terminology for stages of care: acute, chronic, recurrent, and wellness.
As the chiropractic profession delineates its role in the emerging health care marketplace, it will become increasingly important that the scope of appropriate chiropractic care is clearly defined relative to overall patient case management. Therefore, the Council on Chiropractic Guidelines and Practice Parameters engaged in a multidisciplinary consensus process addressing the terminology related to "levels of care." A formal consensus process was conducted in early 2009, following the RAND/UCLA method for rating appropriateness. Panelists were selected to provide a broad representation of the profession in terms of geographic location and organizational affiliation, and an attempt was made to include members of other professions, including representation from third-party payors. The Delphi process was conducted electronically in January-February 2009. A nominal group panel was conducted through an online meeting service using an experienced group facilitator. Twenty-seven panelists were selected; all but 3 were doctors of chiropractic. Six of the panelists had experience as consultants with third-party payors. Fifteen seed statements were circulated to the Delphi panel. Consensus was reached on all statements after 3 Delphi rounds, with further refinements made through the nominal group panel. By using a recognized formal consensus process, the Council on Chiropractic Guidelines and Practice Parameters has endeavored to establish a set of terms that are acceptable to the chiropractic community in order to facilitate their use within the broader health care community.


Comparative radiopacity of flowable resin composites.
This in vitro investigation aimed to determine the radiopacity of eight recently introduced "flowable" restorative resin composites and to compare their radiodensities to those of enamel, dentin, and a widely used composite marketed for universal application in anterior and posterior teeth. A total of 45 composite specimens were fabricated in cylindrical molds and exposed to a curing light for 120 seconds. Two methods were used to determine radiopacity: a transmission densitometer and a computer-assisted densitometric image analysis station. Of the eight flowable resin composites tested, only three exhibited a radiopacity equal to or greater than that of enamel, a widely followed guideline for clinicians' diagnostic ability. The radiopacity of the remaining five materials was not statistically significantly greater than that of dentin. The level of radiopacity of the tested flowable composites was variable; those with low radiodensity should be avoided in Class II restorations, where a clear determination of recurrent caries by the examining clinician could be compromised.


Intra-operative behavioral tasks in awake humans undergoing deep brain stimulation surgery.
Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through implanted electrodes. Deep brain stimulation was first implemented as a therapeutic modality by Benabid et al. in the late 1980s, when he used this technique to stimulate the ventral intermediate nucleus of the thalamus for the treatment of tremor. Currently, the procedure is used to treat patients who fail to respond adequately to medical management for diseases such as Parkinson's, dystonia, and essential tremor. The efficacy of this procedure for the treatment of Parkinson's disease has been demonstrated in well-powered, randomized controlled trials. Presently, the U.S. Food and Drug Administration has approved DBS as a treatment for patients with medically refractory essential tremor, Parkinson's disease, and dystonia. Additionally, DBS is currently being evaluated for the treatment of other psychiatric and neurological disorders, such as obsessive compulsive disorder, major depressive disorder, and epilepsy. DBS has not only been shown to help people by improving their quality of life, it also provides researchers with the unique opportunity to study and understand the human brain. Microelectrode recordings are routinely performed during DBS surgery in order to enhance the precision of anatomical targeting. Firing patterns of individual neurons can therefore be recorded while the subject performs a behavioral task. Early studies using these data focused on descriptive aspects, including firing and burst rates, and frequency modulation. More recent studies have focused on cognitive aspects of behavior in relation to neuronal activity. This article will provide a description of the intra-operative methods used to perform behavioral tasks and record neuronal data with awake patients during DBS cases. Our exposition of the process of acquiring electrophysiological data will illuminate the current scope and limitations of intra-operative human experiments.


Perceptions regarding the shortened dental arch among dental practitioners in the Western Cape Province, South Africa.
This survey was conducted to determine the knowledge of and opinions related to the shortened dental arch (SDA), among dentists in the Western Cape Province, South Africa. The study sample included two consecutive groups, drawn by a process of randomisation from the registered dentist population that included general dentists, specialists, those who had emigrated and retired dentists. A self-administered questionnaire was mailed, e-mailed and/ or faxed to those selected. Reminders were either e-mailed or made by telephone over a period of six months. A final sample of 84 respondents with a mean age of 43 years (SD = 11.9) was obtained. This represented a response rate of 23% (n = 84) from the final working sample (n = 368), derived from the target group (n = 618) originally contacted. All participants completed an informed consent form in which confidentiality was assured. Several respondents (40%) said they had heard about the SDA while at university, which would be in line with the age range of respondents in relation to introduction of the concept into dental curricula. As many as 62% had never read any research articles related to the concept which could partly account for the low response rate. The majority (86%) felt that patients can function with a SDA and that they would recommend acceptance to their patients. Respondents know of the potential benefit that the SDA may have for their patients and see it as a viable alternative treatment option for the partially dentate patient, even though their level of current knowledge of the subject must be considered questionable.


Structural basis of non-specific lipid binding in maize lipid-transfer protein complexes revealed by high-resolution X-ray crystallography.
Non-specific lipid-transfer proteins (nsLTPs) are involved in the movement of phospholipids, glycolipids, fatty acids, and steroids between membranes. Several structures of plant nsLTPs have been determined both by X-ray crystallography and nuclear magnetic resonance. However, the detailed structural basis of the non-specific binding of hydrophobic ligands by nsLTPs is still poorly understood. In order to gain a better understanding of the structural basis of the non-specific binding of hydrophobic ligands by nsLTPs and to investigate the plasticity of the fatty acid binding cavity in nsLTPs, seven high-resolution (between 1.3 A and 1.9 A) crystal structures have been determined. These depict the nsLTP from maize seedlings in complex with an array of fatty acids.A detailed comparison of the structures of maize nsLTP in complex with various ligands reveals a new binding mode in an nsLTP-oleate complex which has not been seen before. Furthermore, in the caprate complex, the ligand binds to the protein cavity in two orientations with equal occupancy. The volume of the hydrophobic cavity in the nsLTP from maize shows some variation depending on the size of the bound ligands. The structural plasticity of the ligand binding cavity and the predominant involvement of non-specific van der Waals interactions with the hydrophobic tail of the ligands provide a structural explanation for the non-specificity of maize nsLTP. The hydrophobic cavity accommodates various ligands from C10 to C18. The C18:1 ricinoleate with its hydroxyl group hydrogen bonding to Ala68 possibly mimics cutin monomer binding which is of biological importance. Some of the myristate binding sites in human serum albumin resemble the maize nsLTP, implying the importance of a helical bundle in accommodating the non-specific binding of fatty acids.


Chitinase-like proteins as regulators of innate immunity and tissue repair: helpful lessons for asthma?
Chitinases and chitinase-like proteins (CLPs) belong to the glycoside hydrolase family 18 of proteins. Chitinases are expressed in mammals and lower organisms, facilitate chitin degradation, and hence act as host-defence enzymes. Gene duplication and loss-of-function mutations of enzymatically active chitinases have resulted in the expression of a diverse range of CLPs across different species. CLPs are genes that are increasingly associated with inflammation and tissue remodelling not only in mammals but also across distant species. While the focus has remained on understanding the functions and expression patterns of CLPs during disease in humans, studies in mouse and lower organisms have revealed important and overlapping roles of the CLP family during physiology, host defence and pathology. This review will summarise recent insights into the regulatory functions of CLPs on innate immune pathways and discuss how these effects are not only important for host defence and tissue injury/repair after pathogen invasion, but also how they have extensive implications for pathological processes involved in diseases such as asthma.


A non-intensive stroke unit reduces functional disability and the need for long-term hospitalization.
In a prospective controlled trial we compared the clinical outcome for unselected acute stroke patients in a non-intensive stroke unit (n = 110) and in general medical wards (n = 183). The patients were comparable in age, marital state and functional impairment on admission. Case fatality rates over the first year after the stroke were similar in the two groups. By three months after the stroke, 15% of the survivors initially admitted to the stroke unit and 39% of those admitted to general medical wards remained hospitalized (p less than 0.001). The corresponding figures by one year after the cerebrovascular accident were 12% and 28%, respectively (p less than 0.05). A greater proportion of surviving stroke unit patients was independent in walking (0.10 greater than p greater than 0.05), personal hygiene (p less than 0.05) and dressing (p less than 0.001). Essential features of the stroke unit are team work headed by a stroke nurse, staff, patient and family education and very early onset of rehabilitation. We conclude that this strategy improves functional outcome and reduces the need for long-term hospital care.


A base-specific recognition signal in the 5' consensus sequence of rotavirus plus-strand RNAs promotes replication of the double-stranded RNA genome segments.
Replication of the segmented double-stranded (ds)RNA genome of rotavirus requires the viral RNA-dependent RNA polymerase (RdRP) to use 11 different (+)RNAs as templates for (-) strand synthesis. Complementary sequences proximal to the 5' and 3' termini are predicted to direct cyclization of the (+)RNAs by forming panhandle structures from which short highly conserved terminal sequences protrude as single-stranded tails. Cell-free replication assays indicate that such structural organization of the 5'- and 3'-ends is required for efficient dsRNA synthesis. Multiple specifically recognized elements exist at the 3'-end that promote dsRNA synthesis including RdRP-recruitment signals and a (-) strand initiation sequence. In contrast to the 3'-end, the role of the 5'-end has been less well defined. In this study, we determined that the 5'-end contains a base-specific recognition signal that plays an important role in the assembly of the RdRP and cofactors into a stable initiation complex for (-) strand synthesis. The 5' recognition signal is associated with the G2 residue of the 5'-consensus sequence, a residue that shows absolute conservation among all rotavirus groups (A, B, and C) examined to date. From our results, we suggest that rotavirus (+)RNA cyclization, although likely initiated by 5'- 3' nucleotide complementarity, may be stabilized by RdRP-dependent bridging. Given that synthesis of the (-) strand on the (+)RNA template will disrupt 5'-3' nucleotide interactions, RdRP-dependent bridging may be the sole mechanism by which the dsRNA product can be held in the necessary cyclized conformation required for efficient multiple rounds of transcription.


Effect of phosphodiesterase-5 inhibition on exercise capacity and clinical status in heart failure with preserved ejection fraction: a randomized clinical trial.
Studies in experimental and human heart failure suggest that phosphodiesterase-5 inhibitors may enhance cardiovascular function and thus exercise capacity in heart failure with preserved ejection fraction (HFPEF). To determine the effect of the phosphodiesterase-5 inhibitor sildenafil compared with placebo on exercise capacity and clinical status in HFPEF. Multicenter, double-blind, placebo-controlled, parallel-group, randomized clinical trial of 216 stable outpatients with HF, ejection fraction ≥50%, elevated N-terminal brain-type natriuretic peptide or elevated invasively measured filling pressures, and reduced exercise capacity. Participants were randomized from October 2008 through February 2012 at 26 centers in North America. Follow-up was through August 30, 2012. Sildenafil (n = 113) or placebo (n = 103) administered orally at 20 mg, 3 times daily for 12 weeks, followed by 60 mg, 3 times daily for 12 weeks. Primary end point was change in peak oxygen consumption after 24 weeks of therapy. Secondary end points included change in 6-minute walk distance and a hierarchical composite clinical status score (range, 1-n, a higher value indicates better status; expected value with no treatment effect, 95) based on time to death, time to cardiovascular or cardiorenal hospitalization, and change in quality of life for participants without cardiovascular or cardiorenal hospitalization at 24 weeks. Median age was 69 years, and 48% of patients were women. At baseline, median peak oxygen consumption (11.7 mL/kg/min) and 6-minute walk distance (308 m) were reduced. The median E/e' (16), left atrial volume index (44 mL/m2), and pulmonary artery systolic pressure (41 mm Hg) were consistent with chronically elevated left ventricular filling pressures. At 24 weeks, median (IQR) changes in peak oxygen consumption (mL/kg/min) in patients who received placebo (-0.20 [IQR, -0.70 to 1.00]) or sildenafil (-0.20 [IQR, -1.70 to 1.11]) were not significantly different (P = .90) in analyses in which patients with missing week-24 data were excluded, and in sensitivity analysis based on intention to treat with multiple imputation for missing values (mean between-group difference, 0.01 mL/kg/min, [95% CI, -0.60 to 0.61]). The mean clinical status rank score was not significantly different at 24 weeks between placebo (95.8) and sildenafil (94.2) (P = .85). Changes in 6-minute walk distance at 24 weeks in patients who received placebo (15.0 m [IQR, -26.0 to 45.0]) or sildenafil (5.0 m [IQR, -37.0 to 55.0]; P = .92) were also not significantly different. Adverse events occurred in 78 placebo patients (76%) and 90 sildenafil patients (80%). Serious adverse events occurred in 16 placebo patients (16%) and 25 sildenafil patients (22%). Among patients with HFPEF, phosphodiesterase-5 inhibition with administration of sildenafil for 24 weeks, compared with placebo, did not result in significant improvement in exercise capacity or clinical status. clinicaltrials.gov Identifier: NCT00763867.


Dendritic receptors designed to bind polyanions in both organic and aqueous media.
This paper reports the synthesis of dendrons containing a spermine unit at their focal point. The dendritic branching is based on l-lysine building blocks, and has terminal oligo(ethyleneglycol) units on the surface. As a consequence of the solubilising surface groups, these dendrons have high solubility in solvents with widely different polarities (e.g., dichloromethane and water). The protonated spermine unit at the focal point is an effective anion binding fragment and, as such, these dendrons are able to bind to polyanions. This paper demonstrates that polyanions can be bound in both dichloromethane (using a dye solubilisation assay) and in water (competitive ATP binding assay). In organic media the dendritic branching appears to have a pro-active effect on the solubilisation of the dye, with more dye being solubilised by higher generations of dendron. On the other hand, in water the degree of branching has no impact on the anion binding process. We propose that in this case, the spermine unit is effectively solvated by the bulk solvent and the dendritic branching does not need to play an active role in assisting solubility. Dendritic effects on anion binding have therefore been elucidated in different solvents. The dendritic branching plays a pro-active role in providing the anion binding unit with good solubility in apolar solvent media.


The economic costs of congenital bilateral permanent childhood hearing impairment.
The objective of this study was to estimate the economic costs of bilateral permanent childhood hearing impairment (PCHI) in the preceding year of life for children aged 7 to 9 years. A cost analysis was conducted by using a birth cohort of children born between 1992 and 1997 in 8 districts of Southern England, of which half had been born into populations exposed to universal newborn screening (UNS). Unit costs were applied to estimates of health, social, and broader resource use made by 120 hearing-impaired children and 63 children in a normally hearing comparison group. Associations between societal costs per child and severity of hearing impairment, language ability score, exposure to UNS, and age of confirmation were analyzed, including adjustment for potential confounders in a linear regression model. The mean societal cost in the preceding year of life at 7 to 9 years of age was 14092.5 pound sterling for children with PCHI, compared with 4206.8 pound sterling for the normally hearing children, a cost difference of 9885.7 pound sterling. After adjusting for severity and other potential confounders in a linear regression model, mean societal costs among children with PCHI were reduced by 2553 pound sterling for each unit increase in the z score for receptive language. Using similar regression models, exposure to a program of UNS was associated with a smaller cost reduction of 2213.2 pound sterling, whereas costs were similar between children whose PCHI was confirmed at <9 or >9 months. CONCLUSIONS. The study provides rigorous evidence of the annual health, social, and broader societal cost of bilateral PCHI in the preceding year of life at 7 to 9 years of age and shows that it is related to its severity and has an inverse relationship with language abilities after adjustment for severity.


Indicators of the quality of general practice care of patients with chronic illness: a step towards the real involvement of patients in the assessment of the quality of care.
To develop a list of indicators of the general practice care of people with chronic illnesses considered important by both patients and practitioners and to identify the indicators that are considered relevant for patient assessment of health care quality. Qualitative study with focus group interviews and a written consensus procedure. General practice in the Netherlands in 1993. 34 patients with chronic illness, mostly members of patient organisations, and 19 general practitioners with expertise in either chronic disease management or experience with patient surveys. Aspects of general practice care considered important for the delivery of good quality care that emerged from focus group interviews; the relevance of evaluations of 41 aspects of care for patients explored through the written consensus procedure. Those aspects of general practice care agreed to be both important and relevant by patients and general practitioners were considered to be suitable indicators for patient assessment of the quality of care. Patients and general practitioners differed to some extent in their assessment of the aspects of care that they considered important for quality. They agreed that most indicators of care that related to the ¿doctor-patient relation¿ and to ¿information and support¿ were relevant and therefore suitable as indicators for patient assessment of health care quality. There was less agreement about the relevance of indicators of ¿medical and technical care,¿ ¿availability and accessibility,¿ and ¿organisation of services.¿ Several indicators of the quality of general practice care of patients with chronic illness were thought to be suitable for the patient assessment of healthcare quality, but other indicators were not, mainly because of reservations by general practitioners. Qualitative methods can contribute to the selection of indicators for assessment of the quality of health care in areas where scientific evidence is limited or where patients' and providers' preferences are particularly important.


A Novel Interaction of the Catalytic Subunit of Protein Phosphatase 2A with the Adaptor Protein CIN85 Suppresses Phosphatase Activity and Facilitates Platelet Outside-in αIIbβ3 Integrin Signaling.
The transduction of signals generated by protein kinases and phosphatases are critical for the ability of integrin αIIbβ3 to support stable platelet adhesion and thrombus formation. Unlike kinases, it remains unclear how serine/threonine phosphatases engage the signaling networks that are initiated following integrin ligation. Because protein-protein interactions form the backbone of signal transduction, we searched for proteins that interact with the catalytic subunit of protein phosphatase 2A (PP2Ac). In a yeast two-hybrid study, we identified a novel interaction between PP2Ac and an adaptor protein CIN85 (Cbl-interacting protein of 85 kDa). Truncation and alanine mutagenesis studies revealed that PP2Ac binds to the P3 block ((396)PAIPPKKPRP(405)) of the proline-rich region in CIN85. The interaction of purified PP2Ac with CIN85 suppressed phosphatase activity. Human embryonal kidney 293 αIIbβ3 cells overexpressing a CIN85 P3 mutant, which cannot support PP2Ac binding, displayed decreased adhesion to immobilized fibrinogen. Platelets contain the ∼85 kDa CIN85 protein along with the PP2Ac-CIN85 complex. A myristylated cell-permeable peptide derived from residues 395-407 of CIN85 protein (P3 peptide) disrupted the platelet PP2Ac-CIN85 complex and decreased αIIbβ3 signaling dependent functions such as platelet spreading on fibrinogen and thrombin-mediated fibrin clot retraction. In a phospho-profiling study P3 peptide treated platelets also displayed decreased phosphorylation of several signaling proteins including Src and GSK3β. Taken together, these data support a role for the novel PP2Ac-CIN85 complex in supporting integrin-dependent platelet function by dampening the phosphatase activity.


Collective effects in epithelial cell death and cell extrusion.
Programmed cell death, notably apoptosis, is an essential guardian of tissue homeostasis and an active contributor of organ shaping. While the regulation of apoptosis has been mostly analysed in the framework of a cell autonomous process, recent works highlighted important collective effects which can tune cell elimination. This is particularly relevant for epithelial cell death, which requires fine coordination with the neighbours in order to maintain tissue sealing during cell expulsion. In this review, we will focus on the recent advances which outline the complex multicellular communications at play during epithelial cell death and cell extrusion. We will first focus on the new unanticipated functions of neighbouring cells during extrusion, discuss the contribution of distant neighbours, and finally highlight the complex feedbacks generated by cell elimination on neighbouring cell death.


Reproductive factors and risk of ovarian cancer: a prospective study.
Associations between reproductive factors and risk of ovarian cancer were examined in a prospective study of 60,565 women in Norway. A total of 445 women were diagnosed as having ovarian cancer during follow-up, from 1961 through 1980. The highest risk was observed among nulliparous women, and the risk decreased significantly with increasing parity. The estimated odds ratio for women with 5 or more births compared with one birth was 0.46, after adjustment for age, urban/rural place of residence and occupational class. Neither age at first or last birth, nor age at menarche or menopause, nor marital status, showed significant associations with ovarian cancer risk after adjustment for parity. The apparent protective effect of high parity was observed for epithelial as well as non-epithelial cancers, and for the separate histological types of epithelial cancer except mucinous cystadenocarcinomas.


Interaction of involuntary post-contraction activity with locomotor movements.
Involuntary post-contraction muscle activity may occur after performing a strong long-lasting (about 30 s) isometric muscle contraction (Kohnstamm phenomenon). Here we examined how this putative excitatory state may interact with a locomotor movement. The subjects stood upright and were asked to oppose a rotational force applied to the pelvis for about 30 s either in the clockwise or in the counterclockwise direction. After that, they were asked to perform various motor tasks with the eyes closed. During quiet standing, we observed an involuntary post-contraction torsion of the trunk. During walking, the post-contraction facilitatory effect of body torsion was not overridden by the voluntary activity, but instead significantly influenced the forward locomotor program such that subjects walked along a curved trajectory in the direction of the preceding torsion. In contrast, we did not observe any rotational component when subjects were asked to step in place. We conclude that the post-contraction rotational aftereffect does not transfer to just any motor task but apparently manifests itself in those movements that incorporate the activated axial muscle synergy or rotational component. We argue that central excitability changes following the voluntary effort may contribute to the phenomenon and highlight the role of tonic influences in fine-tuning of the spinal cord.


Feasibility Evaluation of Myocardial Cannabinoid Type 1 Receptor Imaging in Obesity: A Translational Approach.
The aim of this study was to evaluate the feasibility of targeted imaging of myocardial cannabinoid type 1 receptor (CB1-R) and its potential up-regulation in obese mice with translation to humans using [11C]-OMAR and positron emission tomography (PET)/computed tomography (CT). Activation of myocardial CB1-R by endocannabinoids has been implicated in cardiac dysfunction in diabetic mice. Obesity may lead to an up-regulation of myocardial CB1-R, potentially providing a mechanistic link between obesity and the initiation and/or progression of cardiomyopathy. Binding specificity of [11C]-OMAR to CB1-R was investigated by blocking studies with rimonabant in mice. The heart was harvested from each mouse, and its radioactivity was determined by γ-counter. Furthermore, [11C]-OMAR dynamic micro-PET/CT was carried out in obese and normal-weight mice. Ex vivo validation was performed by droplet digital polymerase chain reaction (absolute quantification) and RNAscope Technology (an in situ ribonucleic acid analysis platform). Subsequently, myocardial CB1-R expression was probed noninvasively with intravenous injection of CB1-R ligand [11C]-OMAR and PET/CT in humans with advanced obesity and normal-weight human control subjects, respectively. Rimonabant significantly blocked OMAR uptake in the heart muscle compared with vehicle, signifying specific binding of OMAR to the CB1-R in the myocardium. The myocardial OMAR retention quantified by micro-PET/CT in mice was significantly higher in obese compared with normal-weight mice. Absolute quantification of CB1-R gene expression with droplet digital polymerase chain reaction and in situ hybridization confirmed CB1-R up-regulation in all major myocardial cell types (e.g., cardiomyocytes, endothelium, vascular smooth muscle cells, and fibroblasts) of obese mice. Obese mice also had elevated myocardial levels of endocannabinoids anandamide and 2-arachidonoylglycerol compared with lean mice. Translation to humans revealed higher myocardial OMAR retention in advanced obesity compared with normal-weight subjects. Noninvasive imaging of cardiac CB1-R expression in obesity is feasible applying [11C]-OMAR and PET/CT. These results may provide a rationale for further clinical testing of CB1-R-targeted molecular imaging in cardiometabolic diseases.


Mining clinical big data for drug safety: Detecting inadequate treatment with a DNA sequence alignment algorithm.
Health data mining can bring valuable information for drug safety activities. We developed a visual analytics tool to find specific clinical event sequences within the data contained in a clinical data warehouse. To this aim, we adapted the Smith-Waterman DNA sequence alignment algorithm to retrieve clinical event sequences with a temporal pattern from the electronic health records included in a clinical data warehouse. A web interface facilitates interactive query specification and result visualization. We describe the adaptation of the Smith-Waterman algorithm, and the implemented user interface. The evaluation with pharmacovigilance use cases involved the detection of inadequate treatment decisions in patient sequences. The precision and recall results (F-measure = 0.87) suggest that our adaptation of the Smith-Waterman-based algorithm is well-suited for this type of pharmacovigilance activities. The user interface allowed the rapid identification of cases of inadequate treatment.


Sinorhizobium meliloti phospholipase C required for lipid remodeling during phosphorus limitation.
Rhizobia are Gram-negative soil bacteria able to establish nitrogen-fixing root nodules with their respective legume host plants. Besides phosphatidylglycerol, cardiolipin, and phosphatidylethanolamine, rhizobial membranes contain phosphatidylcholine (PC) as a major membrane lipid. Under phosphate-limiting conditions of growth, some bacteria replace their membrane phospholipids with lipids lacking phosphorus. In Sinorhizobium meliloti, these phosphorus-free lipids are sulfoquinovosyl diacylglycerol, ornithine-containing lipid, and diacylglyceryl trimethylhomoserine (DGTS). Pulse-chase experiments suggest that the zwitterionic phospholipids phosphatidylethanolamine and PC act as biosynthetic precursors of DGTS under phosphorus-limiting conditions. A S. meliloti mutant, deficient in the predicted phosphatase SMc00171 was unable to degrade PC or to form DGTS in a similar way as the wild type. Cell-free extracts of Escherichia coli, in which SMc00171 had been expressed, convert PC to phosphocholine and diacylglycerol, showing that SMc00171 functions as a phospholipase C. Diacylglycerol , in turn, is the lipid anchor from which biosynthesis is initiated during the formation of the phosphorus-free membrane lipid DGTS. Inorganic phosphate can be liberated from phosphocholine. These data suggest that, in S. meliloti under phosphate-limiting conditions, membrane phospholipids provide a pool for metabolizable inorganic phosphate, which can be used for the synthesis of other essential phosphorus-containing biomolecules. This is an example of an intracellular phospholipase C in a bacterial system; however, the ability to degrade endogenous preexisting membrane phospholipids as a source of phosphorus may be a general property of Gram-negative soil bacteria.


The in vivo mitochondrial two-step maturation of human frataxin.
Deficiency in the nuclear-encoded mitochondrial protein frataxin causes Friedreich ataxia (FRDA), a progressive neurodegenerative disorder associating spinocerebellar ataxia and cardiomyopathy. Although the exact function of frataxin is still a matter of debate, it is widely accepted that frataxin is a mitochondrial iron chaperone involved in iron-sulfur cluster and heme biosynthesis. Frataxin is synthesized as a precursor polypeptide, directed to the mitochondrial matrix where it is proteolytically cleaved by the mitochondrial processing peptidase to the mature form via a processing intermediate. The mature form was initially reported to be encoded by amino acids 56-210 (m(56)-FXN). However, two independent reports have challenged these studies describing two different forms encoded by amino acids 78-210 (m(78)-FXN) and 81-210 (m(81)-FXN). Here, we provide evidence that mature human frataxin corresponds to m(81)-FXN, and can rescue the lethal phenotype of fibroblasts completely deleted for frataxin. Furthermore, our data demonstrate that the migration profile of frataxin depends on the experimental conditions, a behavior which most likely contributed to the confusion concerning the endogenous mature frataxin. Interestingly, we show that m(56)-FXN and m(78)-FXN can be generated when the normal maturation process of frataxin is impaired, although the physiological relevance is not clear. Furthermore, we determine that the d-FXN form, previously reported to be a degradation product, corresponds to m(78)-FXN. Finally, we demonstrate that all frataxin isoforms are generated and localized within the mitochondria. The clear identification of the N-terminus of mature FXN is an important step for designing therapeutic approaches for FRDA based on frataxin replacement.


Feline immunodeficiency virus and feline leukemia virus: frequency and associated factors in cats in northeastern Brazil.
Our aims were to determine the frequencies of feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) in owned and stray cats in the northeastern region of Brazil, ascertain the status of FeLV infection, and investigate potential associated factors among the owned cats. Blood samples from 200 asymptomatic owned cats and 30 stray cats were processed using nested PCR and commercial immunochromatographic tests to diagnose infections. To evaluate the factors associated with FIV and/or FeLV in owned cats, a semi-structured interview was conducted with each owner about the animal's environment, and these data were subjected to unconditional logistic regression. The frequencies for owned cats were 6% (12/200) and 3% (6/200) for FIV and FeLV, respectively. No owned cat was positive for both viruses. Stray cats showed frequencies of 6.66% (2/30) and 0% (0/30) for FIV and FeLV, respectively. Contact with other cats and living in peri-urban areas were considered to be risk factors (P < 0.05) for FIV. We did not identify any factors associated with infections with FeLV. Our results confirm the presence of these two retroviruses in the region under study. Our use of different diagnostic techniques allowed us to determine the frequency of retroviruses in the feline population more accurately, particularly with regard to infections by FeLV, which have complex pathogenesis.


The rhomboid fossa of the clavicle as a sex and age estimator.
The costoclavicular (rhomboid) ligament connects the first rib to the clavicle, stabilizing the pectoral girdle. It produces skeletal traits that may be tubercles, roughened impressions, shallow groove-like fossae, deep fossae, or leave no trace. A pit or depression at this site is often called a "rhomboid fossa." While these markings may appear pathological, they are normal variants of the clavicle. Using a large contemporary sample (N = 344:113 females, 231 males), we evaluated the presence of a rhomboid fossa as a sex and age indicator for unidentified skeletal remains. Logistic regression found significant relationships between the presence of a rhomboid fossa and sex and between presence of a rhomboid fossa and age. Fossae were more common in males (36% left, 31% right) than in females (3% left, 8% right). Posterior probabilities suggest that a fossa on the right clavicle is indicative of a male with 81.7% probability; a fossa on the left is indicative of a male with 92.2% probability. Younger individuals more commonly exhibited rhomboid fossae than older individuals, and the largest fossae were most common in males 20-30 years of age. However, the age effect was not conclusive and must be corroborated by other methods. A test of the sex estimation method on an independent sample (26 males, 23 females) found nine males and only one female with fossae present on the left clavicle. When the costoclavicular attachment exhibits an impression, a tubercle, or leaves no trace, this method cannot be used for sex estimation. When a clavicle exhibits a rhomboid fossa, it is likely from a male. The greater difference in fossa expression between the sexes on the left clavicle makes use of the left bone preferable. This technique can corroborate other sex estimates or provide an estimate for unknown individuals in the absence of other skeletal indicators.


Humoral and cell-mediated immune responses of beef and dairy cattle experimentally infested with Psoroptes ovis.
To compare cellular and humoral immune responses of beef (Belgian White and Blue [BWB]) and dairy (Friesian-Holstein [FH]) cattle to Psoroptes ovis infestation and to determine whether P ovis infestation impaired immune responses to infectious bovine rhinotracheitis virus (IBR) vaccine or an immunogenic protein (keyhole-limpet hemocyanin [KLH]). 19 BWB and 6 FH 1-year-old calves. 2 trials were performed. In each trial, 7 (trial 1) or 6 (trial 2) BWB calves and 3 FH calves were experimentally infested with P ovis and 3 BWB calves were maintained as uninfested controls. Animals were inoculated with KLH and IBR virus vaccine twice; 3 BWB calves in each trial were treated with ivermectin. Serum antibody responses to KLH, IBR virus, and P ovis were measured by use of ELISA. A lymphocyte transformation assay was used to determine nonspecific responses to 3 mitogens and specific lymphocyte reactivity to P ovis antigen. In each trial, 3 BWB and 3 FH calves developed clinical signs of psoroptic mange and mites could be recovered. Infested and control animals developed similar antibody titers to KLH and IBR virus. Antibodies to P ovis were detected early in some infested calves, and this was correlated with a marked cell-mediated immune response. Lymphocyte responsiveness to the 3 mitogens was not significantly different among groups. In these calves, infestation with P ovis induced a marked humoral and cell-mediated immune response. Immunosuppression was not evident.


Use of Coronary Computed Tomographic Angiography Findings to Modify Statin and Aspirin Prescription in Patients With Acute Chest Pain.
Coronary CT angiography (CCTA) is used in patients with low-intermediate chest pain presenting to the emergency department for its reliability in excluding acute coronary syndrome (ACS). However, its influence on medication modification in this setting is unclear. We sought to determine whether knowledge of CCTA-based coronary artery disease (CAD) was associated with change in statin and aspirin prescription. We used the CCTA arm of the Rule Out Myocardial Infarction using Computed Angiographic Tomography II multicenter, randomized control trial (R-II) and comparison cohort from the observational Rule Out Myocardial Infarction using Computed Angiographic Tomography I cohort (R-I). In R-II, subjects were randomly assigned to CCTA to guide decision making, whereas in R-I patients underwent CCTA with results blinded to caregivers and managed according to standard care. Our final cohort consisted of 277 subjects from R-I and 370 from R-II. ACS rate was similar (6.9% vs 6.2% respectively, p = 0.75). For subjects with CCTA-detected obstructive CAD without ACS, initiation of statin was significantly greater after disclosure of CCTA results (0% in R-I vs 20% in R-II, p = 0.009). Conversely, for subjects without CCTA-detected CAD, aspirin prescription was lower with disclosure of CCTA results (16% in R-I vs 4.8% in R-II, p = 0.001). However, only 68% of subjects in R-II with obstructive CAD were discharged on statin and 65% on aspirin. In conclusion, physician knowledge of CCTA results leads to improved alignment of aspirin and statin with the presence and severity of CAD although still many patients with CCTA-detected CAD are not discharged on aspirin or statin. Our findings suggest opportunity for practice improvement when CCTA is performed in the emergency department.


"I tried so many diets, now I want to do it differently" - A single case study on coaching for weight loss.
In this single case study, the author presented an in-depth description and analysis of a coaching intervention with focus on weight loss, conducted over 10 sessions in the course of 17 months. The client was a well-educated woman in her late 30s, who had tried many different forms of dieting over the years-with little and no lasting effect. In his coaching approach, the author went beyond a pure behavioural change model, that is, based on the Health Belief Model, and tried to take a whole-life perspective, where the client learned to link specific events and habits in her work life and everyday life with specific eating habits. In their collaborative practice, coach and coachee initiated changes both in regard to diet, physical activity, and healthy life style, in general. In a theoretical section, the change in understanding with regard to overeating was presented. Finally, an intra-active model-viewing the client as a self-reflective individual-was used as theoretical basis. A narrative analysis of the first session and a cross-session examination was presented to show, analyse, and understand the procedure of the coaching approach. Finally, the voice of the coachee was heard in regard to her personal experiences during the process. The data material was based on audio recordings of selected sessions, notes written by the coach from every session, and final written reflections by the coachee.


Genome-wide insights into introgression and its consequences for genome-wide heterozygosity in the Mytilus species complex across Europe.
The three mussel species comprising the Mytilus complex are widespread across Europe and readily hybridize when they occur in sympatry, resulting in a mosaic of populations with varying genomic backgrounds. Two of these species, M. edulis and M. galloprovincialis, are extensively cultivated across Europe, with annual production exceeding 230,000 tonnes. The third species, M. trossulus, is considered commercially damaging as hybridization with this species results in weaker shells and poor meat quality. We therefore used restriction site associated DNA sequencing to generate high-resolution insights into the structure of the Mytilus complex across Europe and to resolve patterns of introgression. Inferred species distributions were concordant with the results of previous studies based on smaller numbers of genetic markers, with M. edulis and M. galloprovincialis predominating in northern and southern Europe respectively, while introgression between these species was most pronounced in northern France and the Shetland Islands. We also detected traces of M. trossulus ancestry in several northern European populations, especially around the Baltic and in northern Scotland. Finally, genome-wide heterozygosity, whether quantified at the population or individual level, was lowest in M. edulis, intermediate in M. galloprovincialis, and highest in M. trossulus, while introgression was positively associated with heterozygosity in M. edulis but negatively associated with heterozygosity in M. galloprovincialis. Our study will help to inform mussel aquaculture by providing baseline information on the genomic backgrounds of different Mytilus populations across Europe and by elucidating the effects of introgression on genome-wide heterozygosity, which is known to influence commercially important traits such as growth, viability, and fecundity in mussels.


The role of handwashing in prevention of endemic intensive care unit infections.
Handwashing is believed to be the most important means of preventing nosocomial infections. Previous studies of healthcare workers (HCWs) have shown that handwashing practices are poor. No one has shown that handwashing practices can be easily improved and that this prevents endemic (nonepidemic) nosocomial infection. Handwashing and infection rates were studied in two intensive care units (ICUs) of a community teaching hospital. Handwashing rates were monitored secretly throughout the study. After six months of observation, we started interventions to increase handwashing. Handwashing increased gradually, but overall rates before (22.0%) and after (29.9%) interventions were not significantly different (p = .071). Handwashing never occurred before intravenous care, whereas it occurred 67.5% for all other indications (p less than .0001). When questioned, nurses felt they were washing appropriately nearly 90% of the time. Infection rates seemed unrelated to handwashing throughout the study, and no clusters of infection were detected. We conclude that handwashing rates, when measured against arbitrary but reasonable standards, are suboptimal, difficult to change and not closely related to evidence of cross-infection. Further, nurses wash hands selectively, depending on the indication for handwashing, and generally believe they are washing much more frequently than an objective observer believes they are.


Investing in the foundation of sustainable development: pathways to scale up for early childhood development.
Building on long-term benefits of early intervention (Paper 2 of this Series) and increasing commitment to early childhood development (Paper 1 of this Series), scaled up support for the youngest children is essential to improving health, human capital, and wellbeing across the life course. In this third paper, new analyses show that the burden of poor development is higher than estimated, taking into account additional risk factors. National programmes are needed. Greater political prioritisation is core to scale-up, as are policies that afford families time and financial resources to provide nurturing care for young children. Effective and feasible programmes to support early child development are now available. All sectors, particularly education, and social and child protection, must play a role to meet the holistic needs of young children. However, health provides a critical starting point for scaling up, given its reach to pregnant women, families, and young children. Starting at conception, interventions to promote nurturing care can feasibly build on existing health and nutrition services at limited additional cost. Failure to scale up has severe personal and social consequences. Children at elevated risk for compromised development due to stunting and poverty are likely to forgo about a quarter of average adult income per year, and the cost of inaction to gross domestic product can be double what some countries currently spend on health. Services and interventions to support early childhood development are essential to realising the vision of the Sustainable Development Goals.


The impact of mentoring during postgraduate training on doctors' career success.
Although mentoring is perceived as key to a successful and satisfying career in medicine, there is a lack of methodologically sound studies to support this view. This study made use of a longitudinal design to investigate the impact of mentoring during postgraduate specialist training on the career success of doctors. We analysed data pertaining to 326 doctors (172 women, 52.8%; 154 men, 47.2%) from a cohort of medical school graduates participating in the prospective SwissMedCareer Study, assessing personal characteristics, the possession of a mentor, mentoring support provided by the development network, and career success. The impact of personal characteristics on having a mentor was investigated using multiple linear regression analysis. The impacts of having a mentor and of having development network mentoring support on career success were analysed using hierarchical multiple regression analysis. Up to 50% of doctors reported having a mentor. A significant gender difference was found, with fewer female than male doctors having a mentor (40.7% versus 60.4% at the fifth assessment; p ≤ 0.001). Apart from gender, significant predictors of having a mentor were instrumentality (β = 0.24, p ≤ 0.01) and extraprofessional concerns (β = -0.15, p ≤ 0.05). Both having a mentor and having career support from the development network were significant predictors of both objective (β = 0.15, p ≤ 0.01; β = 0.17, p ≤ 0.01) and subjective (β = 0.17, p ≤ 0.01; β = 0.14, p ≤ 0.05) career success, but not of career satisfaction. This study confirmed the positive impact of mentoring on career success in a cohort of Swiss doctors in a longitudinal design. However, female doctors, who are mentored less frequently than male doctors, appear to be disadvantaged in this respect. Formal mentoring programmes could reduce barriers to mentorship and promote the career advancement of female doctors in particular.


In vivo deactivation of peripheral, hepatic, and pancreatic insulin action in man.
The in vivo deactivation of insulin action has been studied in 10 lean, nondiabetic subjects using a modification of the euglycemic glucose clamp technique. Following cessation of 40- and 120-mU/m2/min insulin infusions, the serum insulin levels fell to one-half their initial values (mean +/- SE) of 126 +/- 7 and 350 +/- 14 microunits/ml in 7 +/- 1 and 8 +/- 1 min, respectively. The mean incremental glucose disposal rates (IGDR) fell more slowly following discontinuation of the 40- and 120-mU/m2/min insulin infusions, so that the time required for the IGDRs to fall to one-half their initial values (D50 IGDR) were were 42 +/- 5 and 78 +/- 1 min, respectively. Mean hepatic glucose output was totally suppressed during the 40- and 120-mU/m2/min insulin infusions, remained completely suppressed following cessation of the infusions for 50 and 80 mi, and subsequently returned to basal levels. The times required for the HGOs to return to one-half their basal levels (R50 HGO) were 59 +/- 8 and 119 +/- 6 min, respectively. The times required for insulin action to decrease to one-half the initial values in the periphery (D50 IGDR) and in the liver (R50 HGO) were correlated with the preceding steady-state glucose disposal rates in individual subjects (r = 0.75, P less than 0.001 and r = 0.58, P less than 0.05, respectively). The suppression of endogenous insulin secretion by exogenous insulin infusions was also studied in 4 subjects during a total of 5 euglycemic glucose clamps; the mean basal serum C-peptide level was 0.67 +/- 0.24 pmol/ml before administration of the exogenous insulin, fell to 0.34 +/- 0.17 pmol/ml during the steady-state phase of the study, and remained suppressed throughout the duration of the deactivation phase of the glucose clamp. Residual pancreatic insulin secretory capacity was demonstrated by a rise in the serum C-peptide level to 1.77 +/- 0.50 pmol/ml at 120 min following a standardized meal given at the conclusion of the deactivation phase of the glucose clamp. These results demonstrate that the deactivation of insulin action in the periphery, liver, and pancreas lags behind the disappearance of insulin from the plasma. The mechanisms responsible for this lag in in vivo deactivation are not known for certain, but may include slower clearance of insulin form tissue compartments than form the plasma, the necessity for the target tissues to generate specific deactivation signals, or a slow rate of decay of saturable steps in the cellular activation process.


Expanding application of digital pathology in Japan--from education, telepathology to autodiagnosis.
Digital pathology, i.e., applications of digital information technologies to pathology practice, has been expanding in the recent decades and the mode of pathology diagnostic practice is changing with enhanced precision. In the present study the changing processes of digital pathology in Japan were investigated and trends to future were discussed. The changing status of digital pathology was investigated through reviewing the records of annual meetings of the Japanese Research Society of Telepathology and Pathology Informatics (JRST-PI) and of the Japanese pathology related medical and informatics journals. The results of the Japanese questionnaire survey conducted in 2008-2009 on telepathology and virtual slide were also reviewed. In addition effectiveness of an experimental automatic pathology diagnostic aid system using computer artificial intelligence was investigated by checking its rate of correct diagnosis for given prostate carcinoma digital images. Telepathology played a central role in the development of digital pathology in Japan. Both macroscopic and microscopic pathology digital images were routinely generated and used for diagnostic purposes in major hospitals. Virtual slide (VS) digital images were used first for education then for conference, consultation and also gradually for routine diagnosis and telepathology. The experimental automatic diagnostic aid system achieved the rate of correct diagnosis around 95% for prostate carcinoma and its use for automatic mapping of cancerous areas in a given tissue image was successful. Advance in the digital information technologies gave revolutionary impacts on pathology education, conference, consultation, diagnosis, telepathology and also on pathology diagnostic procedures in Japan. The future will be bright for pathologists by the advanced digital pathology but we should pay attention to make the technologies and their effects under our control.


Year-End Reindeer Dreams.
Updated several times a week with posts by a wide variety of authors, AJN's blog Off the Charts allows us to provide more timely-and often more personal-perspectives on professional, policy, and clinical issues. Best of the Blog will be a regular column to bring AJN readers recent posts that we think deserve a wider audience. To read more, please visit: www.ajnoffthecharts.com.


Recent advances in the feeding and nutrition of dairy goats.
There have been recent advances concerning research of the feeding and nutrition of dairy goats in a wide array of areas. Ruminally emitted methane and supplementary feedstuffs to a lesser extent make appreciable contributions to the carbon footprint of dairy goats, with the former affected by type of production system and associated dietary characteristics. Unique behavior of goats necessitates careful consideration of the nature of confinement facilities to achieve optimal production by animals differing in social hierarchy. Physiological conditions such as nutritional needs and perhaps health status may influence diet selection by goats in both grazing and confinement settings. Some research suggests that low concentrations of protein and fat in milk of high-yielding dairy goat breeds could involve the type and nature of dietary ingredients as influencing end products of ruminal fermentation. With the relationship between milk urea nitrogen concentration and efficiency of dietary protein utilization, through future research the measure may be a useful tool for diet formulation as in dairy cattle. Effects of dietary inclusion of sources of fats and oils vary considerably depending on their nature, as is also true for byproduct feedstuffs and conventional ones being substituted for. Supplementation of dairy goats with sources of polyunsaturated fatty acids can affect oxidative stress and various feedstuffs influence antioxidant status; however, research addressing the significance of such changes under practical production settings would be beneficial.


alpha-Thalassaemia.
The large number of naturally occurring mutants of this well-characterized locus provides an excellent opportunity for elucidating the relationship between its structure and function. Comparisons of what has been learned about the alpha-globin locus with complementary observations on the beta-globin locus, provide a strategy for understanding the co-ordinate regulation of eukaryotic gene expression. From a practical point of view it is important to remember that millions of individuals throughout the world are carriers of alpha-thalassaemia and every year many thousands of pregnancies are at risk of producing children with the severe alpha-thalassaemia syndromes. The data summarized here provide the basis for accurately predicting the genotype in such cases and thus enabling appropriate prenatal testing. However, because this is a genetic disease that predominantly affects individuals from countries with limited health resources, simpler and cheaper methods of screening and diagnosis will have to be developed before this information has a significant impact on the attendant morbidity and mortality (see Chapter 9, this volume).


Regional Adoption of Primary Care-Mental Health Integration in Veterans Health Administration Patient-Centered Medical Homes.
Behavioral health integration is important, yet difficult to implement, in patient-centered medical homes. The Veterans Health Administration (VA) mandated evidence-based collaborative care models through Primary Care-Mental Health Integration (PC-MHI) in large PC clinics. This study characterized PC-MHI programs among all PC clinics, including small sites exempt from program implementation, in one VA region. Researchers administered a cross-sectional key informant organizational survey on PC-MHI among VA PC clinics in Southern California, Arizona, and New Mexico (n = 69 distinct sites) from February to May 2018. Researchers analyzed PC clinic leaders' responses to five items about organizational structure and practice management. Researchers received surveys from 65 clinics (94% response rate). Although only 38% were required to implement on-site PC-MHI programs, 95% of participating clinics reported providing access to such services. The majority reported having integrated, colocated, or tele-MH providers (94%) and care management (77%). Most stated same-day services (59%) and "warm" handoffs (56%) were always available, the former varying significantly based on clinic size and distance from affiliated VA hospitals. Regional adoption of PC-MHI was high, including telemedicine, among VA patient-centered medical homes, regardless of whether implementation was required. Small, remote PC clinics that voluntarily provide PC-MHI services may need more support.


Immunofluorescent visualization of mouse interneuron subtypes.
The activity of excitatory neurons is controlled by a highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus, these are divided into 21 different subtypes of GABAergic interneurons based on their expression of different markers, morphology and their electrophysiological properties. Ideally, all can be marked using an antibody directed against the inhibitory neurotransmitter GABA, but parvalbumin, calbindin, somatostatin, and calretinin are also commonly used as markers to narrow down the specific interneuron subtype. Here, we describe a journey to find the necessary immunological reagents for studying GABAergic interneurons of the mouse hippocampus. Based on web searches there are several hundreds of different antibodies on the market directed against these four markers. Searches in the literature databases allowed us to narrow it down to a subset of antibodies most commonly used in publications. However, in our hands the most cited ones did not work for immunofluorescence stainings of formaldehyde fixed tissue sections and cultured hippocampal neurons, and we had to immunostain our way through thirteen different commercial antibodies before finally finding a suitable antibody for each of the four markers. The antibodies were evaluated based on signal-to-noise ratios as well as if positive cells were found in layers of the hippocampus where they have previously been described. Additionally, the antibodies were also tested on sections from mouse spinal cord with similar criteria for specificity of the antibodies. Using the antibodies with a high rating on pAbmAbs, an antibody review database, stainings with high signal-to-noise ratios and location of the immunostained cells in accordance with the literature could be obtained, making these antibodies suitable choices for studying the GABAergic system.


Cardiomyocyte GTP cyclohydrolase 1 and tetrahydrobiopterin increase NOS1 activity and accelerate myocardial relaxation.
Tetrahydrobiopterin (BH4) is an essential cofactor of nitric oxide synthases (NOS). Oral BH4 supplementation preserves cardiac function in animal models of cardiac disease; however, the mechanisms underlying these findings are not completely understood. To study the effect of myocardial transgenic overexpression of the rate-limiting enzyme in BH4 biosynthesis, GTP cyclohydrolase 1 (GCH1), on NOS activity, myocardial function, and Ca2+ handling. GCH1overexpression significantly increased the biopterins level in left ventricular (LV) myocytes but not in the nonmyocyte component of the LV myocardium or in plasma. The ratio between BH4 and its oxidized products was lower in mGCH1-Tg, indicating that a large proportion of the myocardial biopterin pool was oxidized; nevertheless, myocardial NOS1 activity was increased in mGCH1-Tg, and superoxide release was significantly reduced. Isolated hearts and field-stimulated LV myocytes (3 Hz, 35°C) overexpressing GCH1 showed a faster relaxation and a PKA-mediated increase in the PLB Ser16 phosphorylated fraction and in the rate of decay of the [Ca2+]i transient. RyR2 S-nitrosylation and diastolic Ca2+ leak were larger in mGCH1-Tg and ICa density was lower; nevertheless the amplitude of the [Ca2+]i transient and contraction did not differ between genotypes, because of an increase in the SR fractional release of Ca2+ in mGCH1-Tg myocytes. Xanthine oxidoreductase inhibition abolished the difference in superoxide production but did not affect myocardial function in either group. By contrast, NOS1 inhibition abolished the differences in ICa density, Ser16 PLB phosphorylation, [Ca2+]i decay, and myocardial relaxation between genotypes. Myocardial GCH1 activity and intracellular BH4 are a limiting factor for constitutive NOS1 and SERCA2A activity in the healthy myocardium. Our findings suggest that GCH1 may be a valuable target for the treatment of LV diastolic dysfunction.


The paired-object affordance effect.
We demonstrate that right-handed participants make speeded classification responses to pairs of objects that appear in standard co-locations for right-handed actions relative to when they appear in reflected locations. These effects are greater when participants "weight" information for action when deciding if 2 objects are typically used together, compared with deciding if objects typically occur in a given context. The effects are enhanced, and affect both types of decision, when an agent is shown holding the objects. However, the effects are eliminated when the objects are not viewed from the first-person perspective and when words are presented rather than objects. The data suggest that (a) participants are sensitive to whether objects are positioned correctly for their own actions, (b) the position information is coded within an egocentric reference frame, (c) the critical representation involved is visual and not semantic, and (d) the effects are enhanced by a sense of agency. The results can be interpreted within a dual-route framework for action retrieval in which a direct visual route is influenced by affordances for action.


Gopher mound soil reduces growth and affects ion uptake of two annual grassland species.
Portions of an annual serpentine grassland community in California are subject to frequent gopher mound formation. Consequently, studies were undertaken to characterize the effects of mound soils on plant growth and ion uptake. For two of the dominant annual species (Bromus mollis L. and Plantago erecta Morris), growth was reduced in gopher mound soil relative to that in inter-mound soil. A similar reduction in growth was found for plants grown in soils collected at a depth corresponding to the depth of gopher burrowing. This reduction in growth was associated with lower total P and N contents of the soil which were reflected in lower shoot contents of N and P. Additional experiments, however, showed that reduced N and P availabilities in mound soil were not entirely responsible for the growth reduction. Similarly, shoot Ca/Mg ratios were reduced in mound soil but additions of Ca improved the Ca/Mg ratio without improving growth. Growth reductions were associated with altered shoot concentrations of microelements, particularly elevated levels of Mn. A competition experiment between Plantago and Bromus showed that Bromus was more competitive than Plantago in mound and inter-mound soils and that soil type had only small affects on the nature of the interaction between the two species.


Cardiorespiratory fitness in children: Evidence for criterion-referenced cut-points.
Criterion-referenced cut-points for field-based cardiorespiratory fitness for children (CRF) are lacking. This study determined: (a) the association between CRF and obesity, (b) the optimal cut-points for low CRF associated with obesity in children, and (c) the association between obesity and peak oxygen uptake ([Formula: see text]) estimated from the 20-m shuttle run test using two different prediction equations. A total of 8,740 children aged 10.1±1.2 were recruited from 11 sites across Canada. CRF was assessed using 20mSRT reported as running speed at the last completed stage, number of completed laps and predicted [Formula: see text], which was estimated at the age by sex level using the Léger et al. and FitnessGram equations. Body mass index and waist circumference z-scores were used to identify obesity. Receiver operating characteristic (ROC) curves and logistic regression determined the discriminatory ability of CRF for predicting obesity. 20mSRT had satisfactory predictive ability to detect obesity estimated by BMI, WC, and BMI and WC combined (area under the curve [AUC]>0.65). The FitnessGram equation (AUC>0.71) presented somewhat higher discriminatory power for obesity than the equation of Léger et al. (AUC>0.67) at most ages. Sensitivity was strong (>70%) for all age- and sex-specific cut-points, with optimal cut-points in 8- to 12-year-olds for obesity identified as 39 mL•kg-1•min-1 (laps: 15; speed: 9.0 km/h) and 41 mL•kg-1•min-1 (laps: 15-17; speed: 9.0 km/h) for girls and boys, respectively. 20mSRT performance is negatively associated with obesity and CRF cut-points from ROC analyses have good discriminatory power for obesity.


Effects of whey isolate, creatine, and resistance training on muscle hypertrophy.
Studies that have attributed gains in lean body mass to dietary supplementation during resistance exercise (RE) training have not reported these changes alongside adaptations at the cellular and subcellular levels. Therefore, the purpose of this study was to examine the effects of two popular supplements--whey protein (WP) and creatine monohydrate (CrM) (both separately and in combination)--on body composition, muscle strength, fiber-specific hypertrophy (i.e., type I, IIa, IIx), and contractile protein accrual during RE training. In a double-blind randomized protocol, resistance-trained males were matched for strength and placed into one of four groups: creatine/carbohydrate (CrCHO), creatine/whey protein (CrWP), WP only, or carbohydrate only (CHO) (1.5 g x kg(-1) body weight per day). All assessments were completed the week before and after an 11-wk structured, supervised RE program. Assessments included strength (1RM, three exercises), body composition (DEXA), and vastus lateralis muscle biopsies for determination of muscle fiber type (I, IIa, IIx), cross-sectional area (CSA), contractile protein, and creatine (Cr) content. Supplementation with CrCHO, WP, and CrWP resulted in significantly greater (P < 0.05) 1RM strength improvements (three of three assessments) and muscle hypertrophy compared with CHO. Up to 76% of the strength improvements in the squat could be attributed to hypertrophy of muscle involved in this exercise. However, the hypertrophy responses within these groups varied at the three levels assessed (i.e., changes in lean mass, fiber-specific hypertrophy, and contractile protein content). Although WP and/or CrM seem to promote greater strength gains and muscle morphology during RE training, the hypertrophy responses within the groups varied. These differences in skeletal muscle morphology may have important implications for various populations and, therefore, warrant further investigation.


Rapid and portable electrochemical quantification of phosphorus.
Phosphorus is one of the key indicators of eutrophication levels in natural waters where it exists mainly as dissolved phosphorus. Various analytical protocols exist to provide an offsite analysis, and a point of site analysis is required. The current standard method recommended by the Environmental Protection Agency (EPA) for the detection of total phosphorus is colorimetric and based upon the color of a phosphomolybdate complex formed as a result of the reaction between orthophosphates and molybdates ions where ascorbic acid and antimony potassium tartrate are added and serve as reducing agents. Prior to the measurements, all forms of phosphorus are converted into orthophosphates via sample digestion (heating and acidifying). The work presented here details an electrochemical adaptation of this EPA recommended colorimetric approach for the measurement of dissolved phosphorus in water samples using screen-printed graphite macroelectrodes for the first time. This novel indirect electrochemical sensing protocol allows the determination of orthophosphates over the range from 0.5 to 20 μg L(-1) in ideal pH 1 solutions utilizing cyclic voltammetry with a limit of detection (3σ) found to correspond to 0.3 μg L(-1) of phosphorus. The reaction time and influence of foreign ions (potential interferents) upon this electroanalytical protocol was also investigated, where it was found that a reaction time of 5 min, which is essential in the standard colorimetric approach, is not required in the new proposed electrochemically adapted protocol. The proposed electrochemical method was independently validated through the quantification of orthophosphates and total dissolved phosphorus in polluted water samples (canal water samples) with ion chromatography and ICP-OES, respectively. This novel electrochemical protocol exhibits advantages over the established EPA recommended colorimetric determination for total phosphorus with lower detection limits and shorter experimental times. Additionally this electrochemical adaptation allows the determination of dissolved phosphorus without the use of ascorbic acid and antimony potassium tartrate as reducing agents (as used in the colorimetric method). The potential portability of this protocol is demonstrated in the development of the PhosQuant electrochemical device and provides a portable device for the rapid electrochemical detection of dissolved phosphorus using screen-printed electrodes.


Functional interaction of Parkinson's disease-associated LRRK2 with members of the dynamin GTPase superfamily.
Mutations in LRRK2 cause autosomal dominant Parkinson's disease (PD). LRRK2 encodes a multi-domain protein containing GTPase and kinase domains, and putative protein-protein interaction domains. Familial PD mutations alter the GTPase and kinase activity of LRRK2 in vitro. LRRK2 is suggested to regulate a number of cellular pathways although the underlying mechanisms are poorly understood. To explore such mechanisms, it has proved informative to identify LRRK2-interacting proteins, some of which serve as LRRK2 kinase substrates. Here, we identify common interactions of LRRK2 with members of the dynamin GTPase superfamily. LRRK2 interacts with dynamin 1-3 that mediate membrane scission in clathrin-mediated endocytosis and with dynamin-related proteins that mediate mitochondrial fission (Drp1) and fusion (mitofusins and OPA1). LRRK2 partially co-localizes with endosomal dynamin-1 or with mitofusins and OPA1 at mitochondrial membranes. The subcellular distribution and oligomeric complexes of dynamin GTPases are not altered by modulating LRRK2 in mouse brain, whereas mature OPA1 levels are reduced in G2019S PD brains. LRRK2 enhances mitofusin-1 GTP binding, whereas dynamin-1 and OPA1 serve as modest substrates of LRRK2-mediated phosphorylation in vitro. While dynamin GTPase orthologs are not required for LRRK2-induced toxicity in yeast, LRRK2 functionally interacts with dynamin-1 and mitofusin-1 in cultured neurons. LRRK2 attenuates neurite shortening induced by dynamin-1 by reducing its levels, whereas LRRK2 rescues impaired neurite outgrowth induced by mitofusin-1 potentially by reversing excessive mitochondrial fusion. Our study elucidates novel functional interactions of LRRK2 with dynamin-superfamily GTPases that implicate LRRK2 in the regulation of membrane dynamics important for endocytosis and mitochondrial morphology.


Modulation of breathing behavior of layered coordination polymers via a solid solution approach: the influence of metal ions on sorption behavior.
Four iso-structural layered coordination polymers (CPs) of Cd(II) and Zn(II) were shown to exhibit breathing transitions with N2/H2 gases. Furthermore, the solid solutions containing Zn(II)/Cd(II) have shown better uptake capacities of gas than parent CPs. These results give some insights into the role of metal ions and anions for fine-tuning the porosity of CPs.


Misincorporation of free m-tyrosine into cellular proteins: a potential cytotoxic mechanism for oxidized amino acids.
In vitro studies demonstrate that the hydroxyl radical converts L-phenylalanine into m-tyrosine, an unnatural isomer of L-tyrosine. Quantification of m-tyrosine has been widely used as an index of oxidative damage in tissue proteins. However, the possibility that m-tyrosine might be generated oxidatively from free L-phenylalanine that could subsequently be incorporated into proteins as an L-tyrosine analogue has received little attention. In the present study, we demonstrate that free m-tyrosine is toxic to cultured CHO (Chinese-hamster ovary) cells. We readily detected radiolabelled material in proteins isolated from CHO cells that had been incubated with m-[14C]tyrosine, suggesting that the oxygenated amino acid was taken up and incorporated into cellular proteins. m-Tyrosine was detected by co-elution with authentic material on HPLC and by tandem mass spectrometric analysis in acid hydrolysates of proteins isolated from CHO cells exposed to m-tyrosine, indicating that free m-tyrosine was incorporated intact rather than being metabolized to other products that were subsequently incorporated into proteins. Incorporation of m-tyrosine into cellular proteins was sensitive to inhibition by cycloheximide, suggesting that protein synthesis was involved. Protein synthesis using a cell-free transcription/translation system showed that m-tyrosine was incorporated into proteins in vitro by a mechanism that may involve L-phenylalanine-tRNA synthetase. Collectively, these observations indicate that m-tyrosine is toxic to cells by a pathway that may involve incorporation of the oxidized amino acid into proteins. Thus misincorporation of free oxidized amino acids during protein synthesis may represent an alternative mechanism for oxidative stress and tissue injury during aging and disease.


Anesthetic considerations of percutaneous transcatheter aortic valve implantation: first attempt in Korea -A report of 2 cases-.
Conventional aortic valve replacement for severe aortic stenosis is associated with a high operative mortality in the elderly patients with significant comorbidities, including severe respiratory dysfunction, renal insufficiency, and compromised cardiac function. Human transcatheter aortic valve implantation was first reported in 2002 and has become a valid alternative in selected high-risk patients in Europe and North America. This article describes the first attempt of transfemoral transcatheter aortic valve implantation in Korea. The procedure was applied in two consecutive patients with severe aortic stenosis. Despite several intra-operative complications during procedure, the post-operative outcomes were good for both patients. At post-operative 30 days there was satisfactory prosthetic valve function and hemodynamic stability.


Regulation of T-cell chemotaxis by programmed death-ligand 1 (PD-L1) in dry eye-associated corneal inflammation.
PURPOSE. Given that dry eye disease (DED) is associated with T cell-mediated inflammation of the ocular surface and that PD-L1 is an important negative or inhibitory regulator of immune responses constitutively expressed at high levels by corneal epithelial cells, the authors studied the expression and function of PD-L1 in DED. METHODS. Dry eye was induced in untreated wild-type mice, PD-L1(-/-) mice, and wild-type mice treated with anti-PD-L1 antibody by exposing these mice to a desiccating environment in the controlled environment chamber modified with subcutaneous administration of scopolamine. Real-time PCR was used to quantify the expression of chemokine gene transcript levels of multiple CC and CXC chemokine ligands and receptors. Epifluorescence microscopy was used to evaluate corneal infiltration of CD3(+) T cells after immunohistochemical staining. RESULTS. The increased expression of specific chemokine ligands and receptors in PD-L1(-/-) corneas of normal mice is associated with significant increases in T-cell homing into these corneas. Similar, and more enhanced, increases in T-cell infiltration were observed in PD-L1(-/-) DED mice or DED mice treated with anti-PD-L1 antibody compared with controls. In addition, the authors found significantly decreased expression of PD-L1 by corneal epithelial cells in DED and significantly increased corneal fluorescein staining score with PD-L1 functional blockade using anti-PD-L1 antibody. CONCLUSIONS. Downregulation of corneal epithelial PD-L1 amplifies dry eye-associated corneal inflammation and epitheliopathy by increasing the expression of chemokine ligands and receptors that promote T-cell homing to the ocular surface.